5-Aza-CdR对人食管癌细胞株生物学行为及TFPI-2 mRNA表达的影响  

Effects of 5-Aza-2-deoxycytidine on biological behavior of esophageal carcinoma cell line and TFPI-2 mRNA expression

作　　者：杜雅冰[1] 邵应举[1] 樊青霞[1] 孙桢[1] 王琳[1] 赵培荣[1] 

机构地区：[1]郑州大学第一附属医院肿瘤内科,河南郑州450002

出　　处：《西安交通大学学报（医学版）》2011年第3期295-298,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的探讨5-氮杂-2-脱氧胞苷(5-Aza-2-deoxycytidine,5-Aza-CdR)干预对食管鳞癌细胞株Eca9706生长增殖及其组织因子途径抑制物2(tissue factor pathway inhibitor-2,TFPI-2)基因表达的影响。方法选取食管鳞癌细胞株Eca9706,并用不同浓度的5-Aza-CdR处理该细胞株。MTT法检测干预前后细胞增长率的变化,FCM法检测干预前后细胞凋亡率的变化,RT-PCR技术检测干预前后Eca9706细胞TFPI-2基因mRNA的表达,免疫组化检测干预前后Eca9706细胞TFPI-2蛋白的表达。结果食管鳞癌细胞株Eca9706存在TFPI-2基因超甲基化状态,经5-Aza-CdR处理后,超甲基化状态解除,细胞增殖受到抑制,细胞凋亡率明显提高(P<0.05);细胞中TFPI-2蛋白表达明显增强,同时mRNA的表达水平也较处理前明显提高(P<0.05)。结论食管癌细胞系TFPI-2基因超甲基化可抑制其mR-NA表达,当其超甲基化解除后,细胞增殖受到抑制,同时细胞凋亡率相应提高。Objective To explore the effects of 5-Aza-2-deoxycytidine（5-Aza-CdR） intervention on the growth and proliferation of Eca9706 cell line and protein expression of tissue factor pathway inhibitor-2（TFPI-2）.Methods Eca9706 cell line was treated by 5-azaCdR of different concentrations;MTT,flow cytometry,immunohistochemistry and RT-PCR were used to determine the cell growth,apoptosis,and the expression of TFPI-2 gene and its protein.Results Eca9706 cell line had hypermethylation of TFPI-2.After demethylation by 5-Aza-CdR treatment,the proliferation of Eca9706 was inhibited,the apoptosis rate was also increased significantly in the concentration-dependent manner.RT-PCR detected that mRNA expression of TFPI-2 gene in Eca9706 cell line recovered significantly（P〈0.05）.Conclusion 5-Aza-CdR can slow the growth of Eca9706 cell,increase the apoptosis rate,reactivate the TFPI-2 gene transcription and protein expression by demethylation

关 键 词：食管癌 组织因子途径抑制物-2 5-氮杂-2-脱氧胞苷 甲基化 免疫组化 逆转录聚合酶链反应 凋亡 

分 类 号：R730.5[医药卫生—肿瘤]